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Information on EC 2.3.1.8 - phosphate acetyltransferase and Organism(s) Methanosarcina thermophila and UniProt Accession P38503

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EC Tree
     2 Transferases
         2.3 Acyltransferases
             2.3.1 Transferring groups other than aminoacyl groups
                2.3.1.8 phosphate acetyltransferase
IUBMB Comments
Also acts with other short-chain acyl-CoAs.
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This record set is specific for:
Methanosarcina thermophila
UNIPROT: P38503
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Word Map
The taxonomic range for the selected organisms is: Methanosarcina thermophila
The enzyme appears in selected viruses and cellular organisms
Synonyms
phosphotransacetylase, phosphate acetyltransferase, pta-1, pta-2, aywb-pdul, pdul1, pdul2, moth_1181, moth_0864, phosphotransacetylase pta, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetyltransferase, phosphate
-
-
-
-
phosphoacylase
-
-
-
-
phosphotransacetylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + phosphate = CoA + acetyl phosphate
show the reaction diagram
ternary complex kinetic echanism rather than a ping-pong kinetic mechanism. Sustrates bind to the enzyme in a random order
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:phosphate acetyltransferase
Also acts with other short-chain acyl-CoAs.
CAS REGISTRY NUMBER
COMMENTARY hide
9029-91-8
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-phosphate + CoA
acetyl-CoA + phosphate
show the reaction diagram
-
-
-
?
acetyl-CoA + arsenate
CoA + acetyl arsenate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + phosphate
CoA + acetyl phosphate
show the reaction diagram
acetyl-phosphate + CoA
acetyl-CoA + phosphate
show the reaction diagram
-
ternary complex kinetic echanism rather than a ping-pong kinetic mechanism. Sustrates bind to the enzyme in a random order
-
-
r
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + phosphate
CoA + acetyl phosphate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
K+ or NH4+ at concentration above 10 mM required for maximum activity
NH4+
-
above 10 mM
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
-
above 10 mM
2,3-Butanedione
-
almost complete loss of wild-type enzyme activity after 10 min at 10 mM
acetyl-CoA
-
competitive inhibitor versus CoA when acetyl phosphate is at subsaturating levels but it does not inhibit versus CoA when acetyl phosphate is at saturating levels. Acetyl-CoA is a competitive inhibitor versus acetyl phosphate when CoA is at subsaturating levels but it does not inhibit versus acetyl phosphate when CoA is at saturating levels
desulfo-CoA
N-ethylmaleimide
-
-
Phenylglyoxal
-
33% inhibition after preincubation with phenylglyoxal
phosphate
potassium phosphate
-
above 10 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.094 - 22.5
acetyl phosphate
0.037 - 0.185
CoA
0.043 - 1.32
acetyl phosphate
0.034 - 1.694
CoA
0.8 - 6
desulfo-CoA
0.742
phosphate
-
25°C, pH 7.2
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.1 - 5190
CoA
1.32 - 5190
acetyl phosphate
4500
acetyl-CoA
-
-
0.283 - 5190
CoA
0.00467 - 4.53
desulfo-CoA
1500
phosphate
-
25°C, pH 7.2
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 0.0028
desulfo-CoA
0.32 - 0.81
phosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.1
-
calculated from the deduced amino acid sequence
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
PTAS_METTE
333
0
35220
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35200
-
calculated from amino acid sequence
43000
-
1 * 43000, SDS-PAGE
52000
-
gel filtration
70400
-
calculated from the deduced amino acid sequence
71000
71300
-
calculated from hydrodynamic radius obtained from dynamic light scattering
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
monomer
-
1 * 43000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method. Crystal structures of phosphotransacetylase in complex with the substrate CoA reveals one CoA (CoA(1)) bound in the proposed active site cleft and an additional CoA (CoA(2)) bound at the periphery of the cleft. The crystal structures indicat that binding of CoA(1) is mediated by a series of hydrogen bonds and extensive van der Waals interactions with the enzyme and that there are fewer of these interactions between CoA(2) and the enzyme
hanging drop vapor diffusion method
-
hanging-drop vapor diffusion method
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D316E
kcat for the reaction of acetyl phosphate and CoA is 2.4fold lower than wild-type value, Km for CoA is 1.1fold higher than wild-type value, Km for acetyl phosphate is 1.4fold lower than wild-type value
R310A
kcat for the reaction of acetyl phosphate and CoA is 22.6fold lower than wild-type value, Km for CoA is 1.8fold higher than wild-type value, Km for acetyl phosphate is 122fold higher than wild-type value
R310K
kcat for the reaction of acetyl phosphate and CoA is 472fold lower than wild-type value, Km for CoA is 1.8fold higher than wild-type value, Km for acetyl phosphate is 2.9fold higher than wild-type value
R310Q
kcat for the reaction of acetyl phosphate and CoA is 75.2fold lower than wild-type value, Km for CoA is 2.8fold higher than wild-type value, Km for acetyl phosphate is 4.2fold higher than wild-type value
S309A
kcat for the reaction of acetyl phosphate and CoA is 358fold lower than wild-type value, Km for CoA is nearly identical to wild-type value, Km for acetyl phosphate is 1.96fold lower than wild-type value
S309C
kcat for the reaction of acetyl phosphate and CoA is 851fold lower than wild-type value, Km for CoA is1.4 fold higher than wild-type value, Km for acetyl phosphate is 1.4fold higher than wild-type value
S309T
kcat for the reaction of acetyl phosphate and CoA is 337fold lower than wild-type value, Km for CoA is 1.8fold lower than wild-type value, Km for acetyl phosphate is nearly identical to wild-type value
C159A
-
Km similar to wild-type enzyme
C159A/C277A/C312A/C325A
-
Km similar to wild-type enzyme
C159S
-
Km similar to wild-type enzyme
C277A
-
Km similar to wild-type enzyme
C277A/C312A/C325A
-
Km similar to wild-type enzyme
C312A
-
Km similar to wild-type enzyme
C325A
-
Km similar to wild-type enzyme
R133A
-
altered kinetic properties, increased Km for CoA
R133E
-
altered kinetic properties, increased Km for CoA
R133K
-
altered kinetic properties, increased Km for CoA
R133Q
R287Q
-
decreased Km for CoA
R28Q
-
increased Km
R310Q
-
decreased Km for CoA
R87A
-
altered kinetic properties, increased Km for CoA
R87E
-
altered kinetic properties, increased Km for CoA
R87K
-
altered kinetic properties, increased Km for CoA
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
-
5 min, stable up to 70°C in absence of additional salts
80
-
5 min, complete inactivation in absence of additional salts
additional information
-
sulfate and phosphate partially protect against heat inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
sulfate and phosphate partially protect against heat inactivation
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
stable to air
-
487526
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
stable for at least 3 h when stored on ice
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expressed in Escherichia coli
-
expressed in soluble form in Escherichia coli BL21-DE3
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Lundie, L.L.; Ferry, J.G.
Activation of acetate by Methanosarcina thermophila. Purification and characterization of phosphotransacetylase
J. Biol. Chem.
264
18392-18396
1989
Methanosarcina thermophila
Manually annotated by BRENDA team
Iyer, P.P.; Ferry, J.G.
Role of arginines in coenzyme A binding and catalysis by the phosphotransacetylase from Methanosarcina thermophila
J. Bacteriol.
183
4244-4250
2001
Methanosarcina thermophila
Manually annotated by BRENDA team
Latimer, M.T.; Ferry, J.G.
Cloning, sequence analysis, and hyperexpression of the genes encoding phosphotransacetylase and acetate kinase from Methanosarcina thermophila
J. Bacteriol.
175
6822-6829
1993
Methanosarcina thermophila, Methanosarcina thermophila TM-1
Manually annotated by BRENDA team
Rasche, M.E.; Smith, K.S.; Ferry, J.G.
Identification of cysteine and arginine residues essential for the phosphotransacetylase from Methanosarcina thermophila
J. Bacteriol.
179
7712-7717
1997
Methanosarcina thermophila
Manually annotated by BRENDA team
Iyer, P.P.; Lawrence, S.H.; Yennawar, H.P.; Ferry, J.G.
Expression, purification, crystallization and preliminary X-ray analysis of phosphotransacetylase from Methanosarcina thermophila
Acta Crystallogr. Sect. D
59
1517-1520
2003
Methanosarcina thermophila
Manually annotated by BRENDA team
Iyer, P.P.; Lawrence, S.H.; Luther, K.B.; Rajashankar, K.R.; Yennawar, H.P.; Ferry, J.G.; Schindelin, H.
Crystal structure of phosphotransacetylase from the methanogenic archaeon Methanosarcina thermophila
Structure
12
559-567
2004
Methanosarcina thermophila
Manually annotated by BRENDA team
Lawrence, S.H.; Luther, K.B.; Schindelin, H.; Ferry, J.G.
Structural and functional studies suggest a catalytic mechanism for the phosphotransacetylase from Methanosarcina thermophila
J. Bacteriol.
188
1143-1154
2006
Methanosarcina thermophila (P38503), Methanosarcina thermophila
Manually annotated by BRENDA team
Lawrence, S.H.; Ferry, J.G.
Steady-state kinetic analysis of phosphotransacetylase from Methanosarcina thermophila
J. Bacteriol.
188
1155-1158
2006
Methanosarcina thermophila
Manually annotated by BRENDA team
Nemeti, B.; Gregus, Z.
Mechanism of thiol-supported arsenate reduction mediated by phosphorolytic-arsenolytic enzymes: I. The role of arsenolysis
Toxicol. Sci.
110
270-281
2009
Clostridium kluyveri, Methanosarcina thermophila
Manually annotated by BRENDA team
Gregus, Z.; Roos, G.; Geerlings, P.; Nemeti, B.
Mechanism of thiol-supported arsenate reduction mediated by phosphorolytic-arsenolytic enzymes: II. Enzymatic formation of arsenylated products susceptible for reduction to arsenite by thiols
Toxicol. Sci.
110
282-292
2009
Methanosarcina thermophila
Manually annotated by BRENDA team